Dear netter:
For conventional ELISA, we sequencially coat antigen and add first
antibody, second antibody, and
then substrate. We can adjust the dilution factors of coating antigen,
first/second antigen. But I think we have to keep that the following
agent should be added sufficiantly to former agent; for example, first
antibody should be sufficiantly added not to remain coated antigen
unbound.
Can it be absolutely true? I think so, but my colleague have different
opinion. I think there is no changes of OD ratio depending on coated
antigen, first antibody, and second antibody, if following agents are
sufficient for binding to former agents. My collegue have the opinion
that OD ratio can be changed depending the amount of coated antigen, and
first/second antibody; sometimes invertion of OD ratio also can be
possible. The supposition that the amount of following agents should be
sufficiently added not to remain coated antigen unbound can be destroyed
.
I think the conditioning of agents can be adjusted within my supposition
mentioned in first paragraph. I appreciated for your comments. Thanks.
sincerely,
--
Park, Seung Kyu, M. Sc.
mailto:skpark at gaebyok.wonkwang.ac.krhttp://210.112.141.58/
Tel/Fax : +82-653-850-6768(lab), +82-653-831-0473(APT)
Dept. Parasitol., Won Kwang Univ., 344-2, Sinyong-dong,
Iksan-si, Chunbuk, 570-749, Korea.